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MSP2292
(Plasmid #70707)

Ordering

Item Catalog # Description Quantity Price (USD)
Plasmid 70707 Standard format: Plasmid sent in bacteria as agar stab 1 $65

This material is available to academics and nonprofits only.

Backbone

  • Vector backbone
    pACYCDuet-1
  • Backbone manufacturer
    EMD Millipore
  • Vector type
    Bacterial Expression, CRISPR

Growth in Bacteria

  • Bacterial Resistance(s)
    Chloramphenicol
  • Growth Temperature
    37°C
  • Growth Strain(s)
    DH5alpha
  • Copy number
    Low Copy

Gene/Insert 1

  • Gene/Insert name
    mammalian codon-optimized KKH variant Staphylococcus aureus Cas9(E782K/N968K/R1015H)-NLS-3xFlag, and SaCas9 sgRNA(84)
  • Species
    S. aureus
  • Mutation
    E782K, N968K and R1015H in SaCas9
  • Promoter T7
  • Tags / Fusion Proteins
    • NLS (C terminal on insert)
    • 3x FLAG (C terminal on insert)

Cloning Information for Gene/Insert 1

  • Cloning method Unknown
  • 5′ sequencing primer ACYCDuetUP1
  • 3′ sequencing primer oBK390 (5'-GCAAATTCGACCCGGTCGTCG-3')
  • (Common Sequencing Primers)

Gene/Insert 2

  • Gene/Insert name
    SaCas9 sgRNA (+84)
  • Promoter T7

Cloning Information for Gene/Insert 2

Resource Information

Depositor Comments

For this plasmid, the sequence of the sgRNA spacer is GTCGCCCTCGAACTTCACCT. The length of the sgRNA (not including the spacer sequence) is 84 nucleotides, where this truncated repeat/anti-repeat sgRNA is similar to that described previously by Ran et al., Nature 2015 (PubMed ID: 25830891) and conveys higher activity than the canonical full length sgRNA.

How to cite this plasmid ( Back to top)

These plasmids were created by your colleagues. Please acknowledge the Principal Investigator, cite the article in which the plasmids were described, and include Addgene in the Materials and Methods of your future publications.

  • For your Materials & Methods section:

    MSP2292 was a gift from Keith Joung (Addgene plasmid # 70707 ; http://n2t.net/addgene:70707 ; RRID:Addgene_70707)
  • For your References section:

    Broadening the targeting range of Staphylococcus aureus CRISPR-Cas9 by modifying PAM recognition. Kleinstiver BP, Prew MS, Tsai SQ, Nguyen NT, Topkar VV, Zheng Z, Joung JK. Nat Biotechnol. 2015 Nov 2. doi: 10.1038/nbt.3404. 10.1038/nbt.3404 PubMed 26524662